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Newcastle disease virus (NDV) causes a highly contagious and devastating disease in poultry. ND causes heavy economic losses to the global poultry industry by decreasing the growth rate, decrease in egg production high morbidity and mortality. Although significant advances have been made in the vaccine development, outbreaks are reported in vaccinated birds. In this study, we report the damage caused by NDV infection in the pancreatic tissues of vaccinated and specific-pathogen-free chickens. The histopathological examination of the pancreas showed severe damage in the form of partial depletion of zymogen granules, acinar cell vacuolization, necrosis, apoptosis, congestion in the large and small vessels, sloughing of epithelial cells of the pancreatic duct, and mild perivascular edema. Increased plasma levels of corticosterone and somatostatin were observed in NDV-infected chicken at three- and five- days post infection (DPI). A slight decrease in the plasma concentrations of insulin was noticed at 5 DPI. Significant changes were not observed in the plasma levels of glucagon. Furthermore, NDV infection decreased the activity and mRNA expression of amylase, lipase, and trypsin from the pancreas. Taken together, our findings highlight that NDV induces extensive tissue damage in the pancreas, decreases the activity and expression of pancreatic enzymes, and increases plasma corticosterone and somatostatin. These findings provide new insights that a defective pancreas may be one of the reasons for decreased growth performance after NDV infection in chickens.

Newcastle disease (ND), which is caused by Newcastle disease virus (NDV), can cause heavy economic losses to the poultry industry worldwide. It is characterised by extensive pathologies of the digestive, respiratory, and nervous systems and can cause severe damage to the reproductive system of egg-laying hens. However, it is unknown whether NDV replicates in the male reproductive system of chickens and induces any pathologies. In this study, we selected a representative strain (i.e. ZJ1) of the most common genotype (i.e. VII) of NDV to investigate whether NDV can induce histological, hormonal, and inflammatory responses in the testes of specific pathogen free (SPF) roosters. NDV infection increased the expression of toll like receptor TLR3, TLR7, MDA5, IFN-α, IFN-β, IFN-γ, IL-8, and CXCLi1 in the testes of NDV-infected roosters at 5 days post-infection (dpi). Severe histological changes, including decrease in the number of Sertoli cells and individualized, shrunken spermatogonia with pyknotic nuclei, were observed at 3 dpi. At 5 dpi, the spermatogenic columns were disorganized, and there were fewer cells, which were replaced by necrotic cells, lipid vacuoles, and proteinaceous homogenous material. A significant decrease in the plasma concentrations of testosterone and luteinizing hormone (LH) and the mRNA expression of their receptors in the testes, steroidogenic acute regulatory protein, cytochrome P450 side-chain cleavage enzyme, and 3β-hydroxysteroid dehydrogenase in the NDV-infected group was observed relative to those in the control group (P < 0.05). Collectively, these results indicate that NDV infection induces a severe inflammatory response and histological changes, which decrease the steroidogenesis.

In Egypt, currently two geographically restricted genotypes of the infectious bronchitis coronavirus (IBV) are circulating with detrimental effects for poultry industry. A sensitive real-time RT-PCR assay targeting the IBV nucleocapsid gene (N) was developed to screen clinical samples for presence of IBV. Conventional RT-PCRs amplifying hypervariable regions (HVRs 1-2 and 3) of the IBV S1 gene were developed and amplificates used for nucleotide sequence-based typing of IBV field strains in Egyptian chickens directly from clinical samples., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Newcastle disease virus (NDV) is a pathogen that most often infects poultry species. In investigating a 2014 outbreak of encephalitis and death among farmed mink (Mustela vison), we found pathological and later experimental evidence that NDV can infect and cause severe encephalitic and pneumonic disease in these animals. Our findings confirm the host range of NDV., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The aim of this study was to investigate the effect of Newcastle disease virus (NDV) on the chicken pancreas. A virulent 9a5b mutant NDV isolate was inoculated intranasally into 32-day-old specific pathogen-free white Leghorn chickens. The pancreas was examined grossly and fixed for histopathological, immunohistochemical and electron microscopical investigations. Inflammatory changes were observed in the peripancreatic tissue at the early stage of infection (12 h post infection) and became more prevalent towards the end of the experiment. Multifocal areas of necrotizing inflammation were detected in the exocrine portion of the pancreas by 5 days post infection (dpi) and became more severe at 10 dpi. The endocrine islets were generally preserved, but slight degenerative changes were observed at 10 dpi. Immunohistochemically, NDV-nucleoprotein (NDV-NP) signals were detected in the peripancreatic tissues (associated with macrophages and other lymphoid cells) by 1 dpi. In the exocrine portion of the pancreas, NDV-NP signals were detected at 5 dpi and increased in intensity and distribution by 10 dpi. NDV particles were confirmed in the cytoplasm of exocrine acinar cells by transmission electron microscopy. CD3-positive cells were observed in the peripancreatic tissues earlier than in the pancreatic tissue. Moreover, in comparison with control chickens, insulin immunoexpression was unchanged, except on the last day of the experiment, when it was slightly reduced. The 9a5b NDV infection induced an inflammatory reaction and viral replication in the peripancreatic tissues earlier than in the pancreatic tissue. Furthermore, necrosis affected mainly the exocrine portion of the pancreas, while the endocrine portion was generally unaffected., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Background: An immune system is a system of biological structures and processes within an organism that protects against diseases by identifying and killing pathogens and tumor cells. Development of novel natural immune enhancers is an urgent issue., Methods: In experiment 1, Newcastle disease antibody titer, levels of IL-6, IL-10 and IFN-γ, and changes in CD3(+), CD4(+) and CD8(+) cells were analyzed. In experiment 2, diarrhea, respiratory symptoms and mortality of broilers were measured., Results: In experiment 1, the results showed that the immune response, such as Newcastle disease antibody and CD4(+) T cells, would be significantly activated by feeding with chitosan and its derivative during 27 days. In experiment 2, the results demonstrated that diarrhea, respiratory symptoms and the mortality of the commercial medicine treatment group were decreased, with infection days increased. However, the mortality of broilers was significantly decreased, while feed intake and weight gain of the birds were significantly increased in the group treated with chitosan and its derivative as compared with other groups., Conclusion: Chitosan and its derivative have strong potential in immune enhancement and disease control, and can be used as a feed additive.

Introduction: The first H5N1 outbreak in Burkina Faso was reported to the World Organization for Animal Health on 3 April 2006. This study aimed to determine the prevalence of avian influenza virus, infectious bronchitis virus, and Newcastle disease virus among domestic and wild birds in highly pathogenic avian influenza (HPAI) H5N1 outbreaks areas., Methodology: We collected paired tracheal and cloacal swabs from 283 birds including 278 domestic and five wild birds (three vultures, one sparrowhawk and one Western Grey Plantain-eater) in the Central Region (Ouagadougou) and the Western Region (Bobo-Dioulasso and Sokoroni) of Burkina Faso. Total RNA extracted from samples were subjected to reverse transcription  and resulting cDNA amplified by PCR using specific primers for detection of Avian Influenza Virus (AIV mainly highly pathogenic H5N1), Infectious Bronchitis Virus (IBV), and Newcastle Disease Virus (NDV) for the first time in Burkina Faso., Results and Conclusions: Our results show that 13.8% (39/283) samples were reactive for NDV, and the prevalence of IBV was 3.9% (11/283). None of the 283 birds were co-infected by AIV, IBV and/or NDV in our study areas. The prevalence of influenza A virus was 3.2% (95% CI: 0-6.6) with a 1.7% (95% CI: 0-3.2) prevalence of H5N1 being detected. Positive cases of H5N1 virus were found in two out of three vultures in Ouagadougou, and in three out of 203 local chickens in the Western Region. These results confirm the presence of influenza A H5N1 virus, IBV and NDV in domestic and wild birds in Burkina Faso.

Avian influenza viruses (AIVs) are important zoonotic pathogens whose natural reservoir is waterfowl. In addition to AIV, waterfowl are often coinfected with other viruses, such as the paramyxoviruses, of which Newcastle disease virus (NDV) is of particular importance because of the highly virulent nature of certain strains of this virus for domestic poultry. In routine surveillance of waterfowl for AIV, a number of cloacal samples were encountered that were positive for AIV by real-time reverse transcription polymerase chain reaction (RT-PCR), but did not yield AIV by inoculation in embryonated chicken eggs. On further testing, these samples were also positive for NDV by conventional RT-PCR. It was hypothesized that if both NDV and AIV are present in a sample, the former may overgrow AIV yielding false-negative AIV results. Such samples were treated with chicken anti-NDV polyclonal antiserum and then inoculated in embryonated chicken eggs. Several samples were found to be positive for different subtypes of AIV, indicating that, in the presence of mixed infection with NDV and AIV, it is imperative to remove the influence of NDV, so a true picture of AIV prevalence emerges. An additional benefit is that information on the circulation of NDV in these birds sheds light on their epidemiologic and ecologic significance.

To investigate the immunosuppressive effects of infectious bursal disease virus (IBDV) and aflatoxin in indigenous chickens of Uganda, Newcastle disease (ND) seronegative chicks were randomly allocated to two treatment groups. Group A chicks were injected intramuscularly at the age of 3 weeks every 2 days up to four times with 0.250 mg aflatoxin B1 per bird, group B was infected occulo-nasally with IBDV 3 days prior to vaccination, while group C was left as a control group. All the chicks from the three groups were then vaccinated with Hitchner B1 vaccine at 21 days of age followed by a secondary vaccination with La Sota vaccine 3 weeks later. Humoral and cell-mediated immune responses were assessed by measuring antibody levels and delayed hypersensitivity reaction post vaccination. Growth performance in the three groups was assessed by weekly body weights while evidence of excretion of vaccinal ND virus was detected by reverse transcription-polymerase chain reaction.A significant (P < 0.05) reduction in the haemagglutination inhibition of ND antibody titre following initial priming with Hitchner B1 and subsequent booster with La Sota vaccines and a delayed hypersensitivity test following sensitization with dinitrochlorobenzene showed aflatoxin to be a more potent immunosuppressant than IBDV. Aflatoxin exerted its maximum effects during primary antibody response in the second and third weeks post vaccination. Aflatoxin and IBDV did not affect growth rates (P > 0.05) but prolonged La Sota vaccine virus excretion in faeces. Under our experimental conditions, aflatoxin and IBDV do not significantly affect the immune response of rural chickens to ND vaccination.

A Newcastle disease virus (NDV) isolated in Mexico and called Chimalhuacan strain was characterised by gene F restriction enzyme analysis and found to be a genotype II velogenic virus. Haematological evaluations and histological studies of bone marrow were conducted on chickens experimentally infected with the Chimalhuacan virus and on control chickens. Within 72 hours post infection (hpi), a 50% decrease in thrombocyte and monocyte counts and a complete cellular depletion in bone marrow islands were evident in the infected group. These findings suggest that the Chimalhuacan strain of NDV causes an early and severe damage of the haematopoietic cells including thrombocyte precursors, which might explain the marked thrombocytopenia detected in early stages of this disease.

The pathogenesis of conjunctivitis caused by Newcastle disease viruses (NDVs) was investigated in 3-week-old specific-pathogen-free chickens. The chickens were inoculated intramuscularly, by eye drop or intranasally, with 10(7) plaque forming units of viscerotropic velogenic Newcastle disease virus (VVNDV), mesogenic NDV or lentogenic NDV. Macroscopically, lower palpebral conjunctivae appeared red with oedematous swelling in chickens inoculated with VVNDV. Histologically, mild lesions were focal hyperplasia of the conjunctival epithelial cells with cellular infiltration in the lamina propria of the conjunctivae in the chickens inoculated with VVNDV. Moderate lesions were vascular necrosis with congestion and haemorrhages, hyperplasia of conjunctival epithelial cells, and oedema and cellular infiltration in the lamina propria of the conjunctivae. Fibrin thrombi were observed in the capillaries of the lamina propria. In marked lesions, the whole conjunctiva was affected with more severe and extensive lesions. The mesogenic and lentogenic NDV strains induced no gross conjunctival lesions, but induced mild conjunctivitis without vascular necrosis.

An experimental pigeon paramyxovirus (pPMV-1) infection was followed by reverse transcription-nested polymerase chain reaction for 31 days after infection, in 16 organs of inoculated or contact pigeons naturally infected with Salmonella Typhimurium. With two exceptions, both groups presented similar results. Typical nervous signs and a green diarrhoea were observed. The spread of pPMV-1 was relatively quick, all organs being largely positive at 4 days after inoculation or contact. The lung, spleen, caecal tonsils, kidneys and brain, for which almost all tested samples remained positive during most of the experiment, seemed to be the principal targets for virus persistence. However, the virus was significantly recovered later in the brain parts and for longer in the trachea of the contact pigeons than of the inoculated ones.

We investigated the interaction between Newcastle disease virus (NDV) and Escherichia coli in cell cultures, embryonated eggs, and 8-wk-old chickens. We measured the interactions on the basis of bacterial adherence and NDV hemagglutination titer in chickens, chicken embryos, and chicken embryo cell culture. Depending on the inoculation order of E. coli, a significant alteration of the growth of NDV was observed in both chickens and chicken embryos. When certain strains of E. coli were given before NDV exposure, the virus titers were lowered. In chickens, the mean virus titer was significantly (P < 0.05) lowered in the crop, the proventriculus, the gizzard, and the jejunum. However, there were no significant differences (P < 0.05) between the two groups for NDV titers in the duodenum, ileum, and cecum. In chicken embryos, when E. coli serotypes O78 and O119:B14 were inoculated before NDV exposure, the mean NDV titers were significantly (P < 0.5) lowered. However, there were no significant differences (P < 0.05) in NDV titer between the two groups when E. coli serotypes O78:K80:NM and O1ab:K NM were inoculated 24 hr before NDV exposure. When NDV was given prior to E. coli exposure, NDV titer was higher in both chickens and chicken embryos. In chickens, when NDV was given 48 hr before E. coli inoculation, NDV was detected in the proventriculus, gizzard, jejunum, ileum, and cecum, whereas no virus was detected in the control groups (NDV only). In the crop, NDV was detected at a significantly (P < 0.05) higher titer in the E. coli-inoculated group when compared with the control group that received NDV alone. In chicken embryos, virus titer was significantly (P < 0.05) higher when NDV was given 24 hr before E. coli inoculation for all three NDV strains used (Ulster and V4 strains). Adherence of E. coli to chicken embryo kidney (CEK) cells was significantly higher (P < 0.05) when the CEK cells were infected first with NDV and then by E. coli. The mean bacterial count per microscopic field in NDV-uninfected monolayers was eight compared with 112 for the NDV-infected monolayers. In approximately 10% of the fields in NDV-infected monolayers, the bacteria were too numerous to count.

Avian pneumoviruses (APVs) are RNA viruses responsible for upper respiratory disease in poultry. Experimental infections are typically less severe than those observed in field cases. Previous studies with APV and Escherichia coli suggest this discrepancy is due to secondary agents. Field observations indicate APV infections are more severe with concurrent infection by Newcastle disease virus (NDV). In the current study, we examined the role of lentogenic NDV in the APV disease process. Two-week-old commercial turkey poults were infected with the Colorado strain of APV. Three days later, these poults received an additional inoculation of either NDV or E. coli. Dual infection of APV with either NDV or E. coli resulted in increased morbidity rates, with poults receiving APV/NDV having the highest morbidity rates and displaying lesions of swollen infraorbital sinuses. These lesions were not present in the single APV, NDV, or E coli groups. These results demonstrate that coinfection with APV and NDV can result in clinical signs and lesions similar to those in field outbreaks of APV.

An adult female bearded vulture (Gypaetus barbatus) in the Tel Aviv University Research Zoo was found dead without previous clinical signs. The predominant pathologic changes were considerable bloody content in the intestines and enlargement of the liver, which had a rubbery consistency with color changes. Microscopic lesions consisted of multifocal histiocytic infiltration in the liver. Newcastle disease virus (NDV) was isolated from a cloacal swab and from the lungs and liver. Intracerebral pathogenicity index of the virus, as estimated in 1-day-old chicks, was repeated three times and had an average value of 1.68, indicating a velogenic strain. Numerous Clostridium septicum bacteria were found on the intestinal surface, but bioassays in which they were orally administered into chickens and mice revealed that, even though they were heavily multiplied in the intestines, they were nonpathogenic. It seems that NDV, documented for the first time in a bearded vulture in Israel, was the likely cause of sudden death.

The interaction between Newcastle disease virus (NDV) and Escherichia coli endotoxin was studied in cell cultures, embryonated chicken eggs, and 8-wk-old chickens. These interactions were evaluated according to the induction of specific or nonspecific resistance in the host system and the virus titer produced in both chicken embryos and chickens. The endotoxin of E. coli induced a decrease in the size of the bursa of Fabricius in live chickens. Escherichia coli endotoxin given intravenously induced plasma antiviral activity in chickens that was interpreted to be interferon, as detected in a vesicular stomatitis virus plaque reduction assay. Endotoxin failed to produced toxic effects in the chicken embryo fibroblasts (CEFs) or to result in any antiviral effect because no change was noted in the number of NDV plaques formed in CEF cultures. When endotoxin was given 3 days before NDV exposure in chickens, the virus titers were significantly (P < 0.05) decreased from a peak of 10(2) to 10(0.18), 10(2.5) to 10(0.18), and 10(2.5) to 0 in the spleens, lungs, and kidneys, respectively, at 72 hr post-NDV inoculation. When endotoxin was given 24 hr after NDV inoculation, the NDV titer significantly (P < 0.05) increased from 10(2.0) to 10(3.5), 10(2.5) to 10(6.5), 10(2.5) to 10(4.5), 0 to 10(2.5) in the spleen, lungs, kidneys, and liver, respectively, at 72 hr after NDV inoculation. In chicken sera, hemagglutination inhibition (HI) titer to NDV was significantly (P < 0.05) enhanced from 1164 to 3127 when endotoxin was given prior to virus inoculation. However, there was a decrease in HI to NDV from 1164 to 727 without a significant difference in chicken sera when NDV was given prior to endotoxin inoculation.

The ameliorative effect of graded levels of dietary sodium bentonite (0.2, 0.4 and 0.6 per cent wt/wt of feed) on in vitro-impaired phagocytosis and suppressed immune response to Newcastle disease vaccine during aflatoxicosis (AF) in broiler chicks was investigated. Both percentage and mean of phagocytic activities were decreased significantly (P < 0.05) in chicks fed 2.5 mg aflatoxin per kg feed. The addition of sodium bentonite was significantly effective in ameliorating the negative effect of AF on the percentage and mean of phagocytosis. The presence of AF alone in the diet depressed the immune response of chicks as measured by haemagglutination inhibition (HI) test. Sodium bentonite was also effective in ameliorating the suppressive effect of AF on the HI -titre in chicks vaccinated against Newcastle disease. The best results obtained when sodium bentonite was added at the rate of 0.4 per cent wt/wt of feed to the AF-containing diets., (Copyright 2000 Harcourt Publishers Ltd.)

A non-virulent Newcastle disease virus (strain APMV-1 96/89 VB) was isolated from a broiler chicken from a backyard flock. Using monoclonal antibodies, the virus was shown to be different from the vaccinal virus strains Hitchner, La Sota and Ulster. The virus was shown to replicate in the pancreas of one-day-old specific pathogen-free chickens infected orally, and the histological lesions observed in the pancreas of chickens inoculated with the fourth chicken passage of the virus five to nine days after infection were consistent with an acute pancreatitis.

Seven 18-wk-old pullets from a commercial layer flock experiencing increased mortality associated with neurologic and respiratory symptoms were submitted to the California Veterinary Diagnostic Laboratory System at the Turlock Branch for necropsy. Clinical signs included depression, torticollis, swollen eyelids, conjunctivitis, and sinusitis. Meningoencephalitis and suppurative inflammation of the cranial air spaces were found on histopathology. The brain, sinuses, and air spaces of the cranium were infected with Pasteurella multocida. Complicating the condition was Mycoplasma gallisepticum infecting the sinus and paramyxovirus-I affecting the trachea.

The effects of a mixed infection of Mycoplasma gallinarum and Newcastle disease virus (F strain) on the tracheal epithelium of village chickens were investigated and observed by scanning electron microscopy. Day-old village chicks were vaccinated intranasally with F strain Newcastle disease virus and inoculated intratracheally on the same day with 10(8) colony forming units of M gallinarum. In another study the chicks were vaccinated and then infected with M gallinarum seven days later. The first group of chicks was euthanased three, seven, 10, 14 and 21 days after vaccination and infection and the vaccinated chicks were euthanased three, seven, 10 and 14 days after infection. In the chicks vaccinated and infected on the same day, major alterations to the tracheal epithelium were visible three days later. There were fewer ciliated cells and the borders of the non-ciliated cells were prominent. Several depressions had formed in the epithelial surface. At higher magnification, clumps of microvilli were visible on some of the non-ciliated cells. Seven days after vaccination and infection, the tracheal epithelium appeared normal, with an increase in the numbers of ciliated cells, although raised borders were observed on the non-ciliated cells in some areas. No clumping of microvilli or depressions in the epithelial surface were observed. In the chicks infected seven days after vaccination, the tracheal epithelium appeared normal with no visible changes on its surface.

Ornithobacterium rhinotracheale was first isolated from broilers in South Africa in 1991. The importance of O. rhinotracheale infections has been established, with growth suppression, respiratory symptoms, and arthritis commonly seen as complications. Dual infection with Newcastle disease and O. rhinotracheale in 28-day-old broilers led to more severe respiratory lesions and higher mortality rates than in birds with only Newcastle disease. It was concluded that the pathogenicity of Newcastle disease virus was enhanced when occurring in combination with O. rhinotracheale. Also of interest was the first isolation of O. rhinotracheale from the liver.

Three isolates of Newcastle disease virus (NDV) from cormorants and turkeys were classified as velogenic neurotropic NDV (VNNDV) by standard pathotyping procedures in specific-pathogen-free white leghorn and white rock chickens. White leghorns inoculated by eye drop, cloaca, intravenous, or intracerebral routes had a higher frequency of nervous signs and mortality than was observed in similarly inoculated white rocks. Two backpassages in white rocks increased the intravenous pathogenicity index of all three isolates, but none had a value after backpassage that was as high as the classical VNNDV strain Texas GB. The breed of chicken used in standard procedures did influence the numerical values of pathotyping determinations, but the differences between breeds were not large enough to change the pathotype assigned to these isolates.

Turkey rhinotracheitis (TRT) virus was first isolated from a commercial broiler chicken with swollen head syndrome (SHS) in Japan. At the same time, Newcastle disease virus (NDV), infectious bronchitis virus (IBV), avian reovirus (ARV), Escherichia coli (E.coli), Morganella morganii, and Proteus mirabilis were also isolated from the same broiler chicken. The presence of antibodies to TRT virus was confirmed in the sera of 34-day-old chickens of the flock with SHS, however the antibodies to TRT virus were undetectable in the sera of 17-day-old chickens. In this investigation, we confirmed avian pneumovirus infection in chickens in Japan, and the virus and other agents may be considered as a cause of SHS.

This study was designed to examine whether vaccine or virulent strains of Newcastle disease virus (NDV) would potentiate the disease caused by Pasteurella anatipestifer infection in turkeys. The studies were conducted in turkeys of two age groups. There were three experiments. In two experiments four-week-old turkeys were exposed either to vaccine or virulent strains of NDV after experimental P anatipestifer infection. In the third experiment 14-week-old turkeys were first exposed to virulent NDV and superimposed with P anatipestifer infection. In experiment 1, one bird died where P anatipestifer was given in combination with the vaccine strain of NDV. However, there was no difference in the clinical signs, gross lesions and histopathology compared with turkeys given P anatipestifer alone. In experiment 3 where turkeys received a virulent strain of NDV in combination with P anatipestifer, birds became dyspnoeic and showed signs of illness. There was a difference in the course of the disease, gross lesions and histopathology when compared with turkeys that received P anatipestifer only.

A severe nervous disease struck fancy pigeons for the first time in Saudi Arabia during February-March, 1992. The morbidity rate was 60% while the case fatality rate was 40%. An avian paramyxovirus-1 was isolated from affected pigeons. The disease was reproduced experimentally in pigeons and the virus was reisolated and identified.

As part of a case-control study designed to identify fowl cholera risk factors, 2087 blood samples were collected from 71 California meat-turkey flocks. Samples were tested for antibodies to three mycoplasmas and four viruses pathogenic for turkeys. Flocks that had antibodies to Newcastle disease virus and/or Mycoplasma meleagridis had an increased risk of having an outbreak of fowl cholera. This information should prove useful for fowl cholera control programs in meat turkeys.

The effect of vitamin A deficiency in the presence or absence of Newcastle disease virus infection (NDV, La Sota strain) on weight of lymphoid organs and on the number and type of circulating white blood cells (WBC) was investigated in chickens. Day-old chickens with limited vitamin A reserves were fed purified diets containing either marginal (ad libitum) or adequate (pair-fed) levels of vitamin A and at 21-28 days of age; half the chickens in each group were infected with NDV. Vitamin A deficiency resulted only in significantly lower absolute and relative weights of bursa of Fabricius and after infection both weights of bursa and thymus were significantly lower. Relative weight of spleen was significantly higher after infection irrespective of vitamin A status. Liver weights were not affected by vitamin A status and/or NDV infection. Both vitamin A deficiency and NDV infection resulted in lymphopenia, while the lowest number of WBC were observed in vitamin A-deficient chickens during the acute phase of NDV (5 days after infection). Subsequent to lymphopenia due to NDV infection, a marked lymphocytosis was observed in controls and to a lesser extent in vitamin A-deficient birds. These results indicate that vitamin A deficiency, which is aggravated by concomitant NDV infection, affects lymphoid cell systems.

The effect of vitamin A deficiency on cytotoxic T lymphocyte (CTL) activity was investigated during the acute phase of disease 7 days after primary inoculation and 1 day after secondary inoculation in chickens with or without Newcastle disease virus (NDV, La Sota strain) infection. Day-old chickens with limited vitamin A reserves were fed purified diets containing either marginal (ad libitum) or adequate (pair-fed) levels of vitamin A, and at 3 weeks of age half of the chickens in each group were infected with NDV. Cytotoxic activity was investigated during the acute phase of disease (7 days after primary inoculation) and 1 day after secondary inoculation, in an assay system with either peripheral blood lymphocytes (PBL) or nonadherent splenocytes as effector cells and adherent splenocytes from the same animal as target cells. After primary inoculation, cytotoxic activity could only be demonstrated in nonadherent splenocytes. Vitamin A deficiency resulted in significantly reduced CTL activity at all effector/target cell ratios tested. After reinfection CTL activity could also be demonstrated in PBL, but only from chickens fed the control diet, suggesting a diminished pool of CTL in vitamin A deficiency. The results of this study indicate that vitamin A deficiency impairs CTL activity - a part of the cell-mediated defense system - and this may have important implications for recovery from viral infection.

Factors playing a part in the development of respiratory disease complex in chickens were investigated in a series of experiments. The experimental infection was produced by exposing chickens to Mycoplasma gallisepticum and the B1 vaccine strain of Newcastle disease virus and later exposing them to aerosols containing the O1:K1 serotype of Escherichia coli. Chickens became susceptible (pericarditis or death) to E. coli 8 days after mixed respiratory disease challenge. One day after respiratory disease challenge, lesions consisted of edema and infiltration with lymphoid cells and heterophils. At the time of susceptibility to E. coli, the lesions were strongly lymphoid with many dense follicular areas and very few heterophils. The incidence of pericarditis and death was similar when the concentration of bacteria in the aerosol inoculum ranged between 10(9)/ml and 10(5)/ml. At the time of maximum susceptibility to aerosol challenge, chickens were less susceptible to intravenously administered E. coli than were the uninfected controls. Resistance of chickens that had been selectively bred for a high (HA) or low (LA) antibody response to sheep erythrocytes was compared. HA chickens were more resistant to respiratory agents and less resistant to E. coli than LA line chickens. When the lines were exposed to respiratory disease followed by exposure to aerosols containing E. coli, the HA line had the lowest incidence of pericarditis and death.

A flock of 4,500 Cobb broilers inoculated with Newcastle disease vaccine intra-ocular strain B1 type at 10 days of age developed clinical signs of the disease 19 days later; the mortality rate was 71%. Necropsy examinations showed characteristic lesions. Newcastle disease virus was isolated and identified in the allantoic fluid of embryonating chicken eggs by haemagglutination and haemagglutination-inhibition tests. Histopathological examination showed that follicles of the bursa were depleted of lymphocytes, had many large cavities and were being repopulated by newly formed healthy lymphocytes. Both the acute and convalescent serum samples were positive for infectious bursal disease antibodies in agar gel precipitation tests. Haemagglutination inhibition titres of the acute and convalescent sera were 20 to 80 and 80 to 640 respectively. The vaccine failure may be due to either the subclinical bursal disease or the highly pathogenic nature of the wild Newcastle disease virus.

Studies on the ocular pathogenesis of Newcastle disease virus (NDV) in rabbits and monkeys revealed that conjunctivitis can only be produced by the virus when conjunctival epithelium is traumatized; multiple instillations of large doses of virus over the intact conjunctiva failed to do so. The presence of serum HI antibodies showed no correlation with the clinical form of the disease in rabbits. However, in monkeys, the disease showed a uniform pattern of immune response irrespective of the strain of the virus. Monkeys may be useful for further studies.

Mixed infections involving mycoplasmas, viruses and bacteria are well recognized in chickens. Synergism has been demonstrated between Mycoplasma gallisepticum and the viruses of Newcastle disease and infectious bronchitis and Escherichia coli, although the outcome of infection is influenced by many factors associated with the host and the organisms. Airsacculitis in broilers due to M. synoviae or M. gallinarum may be precipitated by concurrent respiratory virus infections including vaccine strains. Turkeys, geese and ducks have been less well studied, but similar interactions seem to occur. Such observations may give an indication as to the likely interactions between mycoplasmas, viruses and bacteria in other host species.

The Australian strain of infectious bursal disease virus (IBDV), 002/73, affected the response of chickens to Newcastle disease virus (NDV). The titre of serum antibodies to NDV in chickens infected with IBDV was significantly lower than that of birds infected with NDV alone. It also appeared that IBDV affected NDV excretion from chickens as NDV was more frequently isolated from chickens infected with IBDV, IBDV infection did not alter the pathogenicity of NDV in chickens. This Australian strain of IBDV therefore appeared to be immunodepressive in one-day-old chickens.

Oocysts were recovered from chickens experimentally infected with the Mukteswar strain of Newcastle disease virus and Eimeria acervulina or E tenella. Oocysts sterilised by sodium hypochlorite solution were washed, ruptured and inoculated into embryonating eggs which were examined after five days by the haemagglutination test for virus. All these tests were negative. Virus, initially detected with oocysts separated from faeces, was no longer present after sporulation in 2 per cent potassium bichromate. The B1 strain of virus survived for up to 30 hours in 2 per cent sodium hydroxide and 2 per cent potassium bichromate but was quickly destroyed by sodium hypochlorite (specific gravity 1.075).

The paramyxovirus of Newcastle disease (NDV) impairs the contact sensitivity to oxazolone in CBA/J mice: in vitro treatment with the infectious virus inhibits the passive transfer of contact sensitivity. Experiments performed with three different virus preparations demonstrate that only the infectious virus is able to inhibit the hypersensitivity reaction while treatment of virus by heat or ultraviolet irradiation wipes out its inhibitory activity. These results suggest that I-NDV depresses the response to oxazolone by its action on T immunoblasts which mediate the response to contactant.

All chickens infected by intranasal and conjunctival routes with a neurovirulent strain of Newcastle disease virus (NDV) developed pneumonitis by 4 days after infection. This was followed 6 to 12 days later by the appearance of severe encephalitis in a significant number of survivors. Histologically, the encephalitis was characterized by neuronal degeneration and perivascular inflammation. In addition, a proliferative vasculitis in the molecular layer of the cerebellum was noted after 30 days, and this persisted for at least 200 days. Although CNS signs were absent during the pneumonic stage, significant amounts of virus were present in the brains of all birds at this time, and viral antigens were easily demonstrable in neurons, glial cells, and endothelial cells. However, when the CNS disease became apparent, viral antigens could not be detected in brains and NDV could regularly be recovered only by application of cocultivation techniques. At this time, the agent was selectively present in birds exhibiting neurologic signs. No evidence for immunopathologic processes was obtained, although hemagglutination inhibiting antibody levels to NDV were elevated in birds with CNS disease. Studies of an avirulent strain of NDV that replicated in the CNS but produced no disease provided evidence that the histologic lesions and the neurologic disease were related to virus multiplication in the brain parenchyma. A mechanism of the pathogenesis of NDV encephalitis is proposed and the disease is discussed relative to other paramyxovirus-induced encephalitic syndromes.